LOCUS       sequence-173823-        3623 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Can be used for in situ hybridisation or other purposes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3623)
  AUTHORS   Hall TE, Martel N, Lo HP, Xiong Z, Parton RG
  TITLE     A plasmid library of full-length zebrafish rab proteins for in vivo 
            cell biology.
  JOURNAL   Cell Logist. 2017 Mar 1;7(1):e1301151. doi: 
            10.1080/21592799.2017.1301151. eCollection 2017.
  PUBMED    28396820
REFERENCE   2  (bases 1 to 3623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Logist."; date: "2017-03-1"; volume: "7(1):e1301151. doi"; pages: " 
            10.1080/21592799.2017.1301151. eCollection 2017"
COMMENT     Sequence Label: Rab35a cds in pGem-T Easy
FEATURES             Location/Qualifiers
     source          1..3623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..27)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     218..237
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     450..472
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     464..481
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     465..481
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     488..507
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        488..506
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      541..550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1252..1270)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1253..1270)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1288..1304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1288..1304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1301..1323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1312..1328
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1336..1342,1343..1360,1361..1366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1381..1402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1519..1536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1690..2278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1770..1789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2449..3240,3241..3309))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3072..3091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3310..3414)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3492..3623,1..324))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
       61 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
      121 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      181 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      241 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      301 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
      361 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      421 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      481 tgaattgtaa tacgactcac tatagggcga attgggcccg acgtcgcatg ctcccggccg
      541 ccatggcggc cgcgggaatt cgattctagc agcacttctt cttccgtttg ctgttctgct
      601 tgagtttaat cacatcgttc tgctgctgct gctgctgttt ggccacagac tctttctttg
      661 cccggagaac caactctgtg atgcagttaa acatctcctc cacattgaca ttttctttcg
      721 cacttgtctc gaacagactg atgcccatct gttctgcgaa cttctgtgca tcattcgtct
      781 ccaccacctt agagtttgga tcatcatttt tattgcccac taatattcga cagacgtcat
      841 cacagttctg gtttatttca tgtagccatc gtttaacatt gacaaaggac tcggcgctgg
      901 taacgtcata cactactatc acaccatggg ttcctctgta atatgtagat gtaatcgttc
      961 ggaatcgctc ttgtccggct gtgtcccata tctgcagttt caccttctct ccattaagct
     1021 ccacagtgcg gatcttgaag tcgactccta tagtggtgat gtaactgcct gaaaatgtgt
     1081 tgtcggcgaa acgcagaaga agactgctct ttccaacacc gctgtcaccg atgatcagca
     1141 gcttgaagag gtgatcgtag tcgcgggcca taatcactag tgaattcgcg gccgcctgca
     1201 ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta ttctatagtg
     1261 tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3421 tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3481 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3541 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3601 atagaccgag atagggttga gtg
//
